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Sufficiency for inducible Caspase-9 safety switch in human pluripotent stem cells and disease cells

Gene Therapy volume 27, pages 525–534 (2020)Cite this article

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Abstract

Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) have promising potential for opening new avenues in regenerative medicine. However, since the tumorigenic potential of undifferentiated pluripotent stem cells (PSCs) is a major safety concern for clinical transplantation, inducible Caspase-9 (iC9) is under consideration for use as a fail-safe system. Here, we used targeted gene editing to introduce the iC9 system into human iPSCs, and then interrogated the efficiency of inducible apoptosis with normal iPSCs as well as diseased iPSCs derived from patients with acute myeloid leukemia (AML-iPSCs). The iC9 system induced quick and efficient apoptosis to iPSCs in vitro. More importantly, complete eradication of malignant cells without AML recurrence was shown in disease mouse models by using AML-iPSCs. In parallel, it shed light on several limitations of the iC9 system usage. Our results suggest that careful use of the iC9 system will serve as an important countermeasure against posttransplantation adverse events in stem cell transplantation therapies.

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Fig. 1: Correlation between inducible caspase-9 (iC9) expression and induced apoptosis kinetics.
Fig. 2: Induced apoptosis in AML mouse model.

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Acknowledgements

We thank Miki Ando and Motoo Watanabe (The University of Tokyo) for helpful discussions; Atsushi Miyawaki and Satoshi Iwano (RIKEN) for kindly providing AkaLuc gene and Akalumine-HCl (TokeOni); and Austin Smith and Yasuhiro Takashima (University of Cambridge) for kindly providing PiggyBac donor and transposase plasmids.

Funding

The project was supported in part by a grant from California Institute for Regenerative Medicine (CIRM; LA1_C12-06917, QUEST DISC2-08874), Weston Havens Foundation, and a research fellowship from Japan Society for the Promotion of Science (JSPS).

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Author notes

  1. Masayuki Iwasaki

    Present address: Department of Advanced Health Science, Institute of Laboratory Animals, Tokyo Women’s Medical University, Tokyo, 162-8666, Japan

  2. These authors contributed equally: Toshinobu Nishimura, Haojun Xu, Masayuki Iwasaki

Authors and Affiliations

  1. Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, 94305, USA

    Toshinobu Nishimura, Masayuki Iwasaki, Daiki Karigane, Brandon Saavedra, Yusuke Takahashi, Fabian P. Suchy, Shinichiro Monobe, Renata M. Martin, Michael L. Cleary, Ravindra Majeti & Hiromitsu Nakauchi

  2. Department of Immunology, Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki, 305-8575, Japan

    Haojun Xu & Akira Shibuya

  3. Ph.D. Program in Human Biology, School of Integrative and Global Majors, University of Tsukuba, Tsukuba, Ibaraki, 305-8575, Japan

    Haojun Xu

  4. Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, 305-8562, Japan

    Manami Ohtaka & Mahito Nakanishi

  5. QIMR Berghofer Medical Research Institute, Brisbane, Queensland, 4006, Australia

    Scott R. Burrows

  6. Life Science Center of Survival Dynamics, Tsukuba Advanced Research Alliance (TARA), University of Tsukuba, Tsukuba, Ibaraki, 305-8575, Japan

    Akira Shibuya

  7. Division of Stem Cell Therapy, Distinguished Professor Unit, The Institute of Medical Science, The University of Tokyo, Tokyo, 108-8639, Japan

    Hiromitsu Nakauchi

Authors
  1. Toshinobu Nishimura
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Contributions

Conceptualization: TN; methodology: TN, HX, and MI; investigation: TN, HX, MI, DK, BS, YT, FPS, and SM; resources: RMM, MO, MN, SRB, MLC, and RM; writing—original draft: TN, HX, and HN; writing—review and editing: TN, HX, FPS, and HN; funding acquisition: TN and HN; and supervision: TN, AS, and HN.

Corresponding authors

Correspondence to Toshinobu Nishimura or Hiromitsu Nakauchi.

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Conflict of interest

RM is a co-founder, equity holder, and serves on the Board of Directors of Forty Seven, Inc. HN is a co-founder, member of the scientific advisory board, and shareholder of Megakaryon Corp., iCELL, Inc. and Century Therapeutics, LLC. The remaining authors declare no conflict of interest.

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Nishimura, T., Xu, H., Iwasaki, M. et al. Sufficiency for inducible Caspase-9 safety switch in human pluripotent stem cells and disease cells. Gene Ther 27, 525–534 (2020). https://doi.org/10.1038/s41434-020-0179-z

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